Terunuma H
Biotherapy Institute of Japan (BIJ) , Tokyo, Japan.
J Stem Cells Regen Med. 2012 Nov 26;8(3):205-6. eCollection 2012.
Cancer, the major killer disease of the century requires a multi-pronged approach and among the latest modalities of treatments, Immunotherapy occupies a promising role. Immunotherapy for cancer was first started to be practised in the NIH and cell based immunotherapy for cancer is in practice for the past three decades. ([1, 2]) There are several literatures from various countries on the successful application of cell based Immunotherapies for various solid tumours and haematological malignancies. ([3-8]) Our team's association with immune cells started when I was working on RNA transcriptome analysis to understand the immune system in HIV carriers which in turn required in vitro expansion of human Natural Killer (NK) cells. ([9]) This led to the customization of protocols which has resulted in successful in vitro expansion, activation of NK cells and T cells for Immunotherapy. The purpose of Biotherapy institute of Japan (BIJ) is to support research and clinical application of immune cells like NK cells, γδT cells, αβT cells, Cytotoxic T lymphocytes (CTL) and Dendritic cells (DC) for application as Autologous Immune Enhancement Therapy (AIET) to fight against cancer. AIET using NK cells, CTLs, DCs etc have been administered for more than 5000 patients since 2004 till date by BIJ.
For AIET using NK cells, the process involves separation of lymphocytes from the peripheral blood of the patient followed by selective NK cell expansion using the expansion kit (BINKIT, BIJ, JAPAN) without feeder layers and then infusion of the expanded-activated NK cells. ([10,11]) As reports suggest that the activity of peripheral blood NK cells are lower in cancer patients compared to normal individuals([12]) and as in vitro expansion of NK cells increases the cytotoxic ability 5 to 10 fold, ([13]) the NK cells are expanded in vivo and then infused to the patient in AIET. We are also working on combination immunotherapy using NK cells and CTLs and also NK cells and peptide pulsed DCs. The principle behind combining NK cells and CTLs is a dual advantage approach combining the innate immune system and adaptive immune system wherein the CTLs will kill the MHC expressing cancer cells while NK cells will kill the MHC non-expressing cancer cells also. ([10]) In case of NK cells and DCs, DCs will in turn activate the CTLs thereby giving rise to the dual advantage mentioned above. We have recently suggested that the AIET using expanded NK cells, particularly in combination with monoclonal antibody drugs, may be very useful tool for cancer immunotherapy. ([14].)
In our studies in NOG SCID mice, activated NK cells were shown to reduce the size of breast cancer cells (MDA-MB231) ([15]) and the volume of ascites also inhibiting lung metastasis of pleural effusion lymphoma (PEL) cells. ([16]) In a primary lung adenocarcinoma patient where AIET was administered in combination with Hyperthermia, the CEA values decreased from 4.8 ng/ml to 1.6 ng/ml, the SLX decreased from 30 U/ml to 27 U/ml, the IAP reduced from 300 μg/ml to 221 μg/ml along with resolution of the lung lesions in four months. A 55 year old woman treated for invasive ductal carcinoma of breast presented with metastasis to the lungs. She was then treated with trastuzumab in combination with multiple injections of activated natural killer (NK) cells (at two week intervals) following which the tumor markers decreased. Progression free survival was 10 months. ([11]) Fifty-two patients with advanced cancers in organs like lung, breast, colon, prostate, liver, kidney, ovary etc, refractory to conventional therapy when treated with a combination of hyperthermia and NK cell-and CTL-based immune cell therapy with low-dose chemotherapy, in 18 of 52 patients, objective responses was observed including one complete response (CR) and 17 partial responses (PR) evaluated according to to the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines. Sixteen patients had stable disease (SD), whereas 18 had progressive disease (PD). Disease control rate was 66% including CR, PR and SD. After treatment for six months, the objective responses and disease control rate were 25% and 52%, respectively. There were no adverse effects in any of these patients. ([17].)
Cancer has to be tackled with a multipronged approach and combining NK cell and CTL cell based AIET with conventional modalities of treatments such as Surgery, Chemotherapy and Radiotherapy as well as other modalities like Hyperthermia, Proton Beam therapy and low dose chemotherapy is effective even in advanced cancers which are refractory to conventional therapeutic modalities.
癌症是本世纪主要的致命疾病，需要采取多管齐下的方法，在最新的治疗方式中，免疫疗法占据着有前景的地位。癌症免疫疗法最初是在美国国立卫生研究院开展的，基于细胞的癌症免疫疗法在过去三十年中一直在实践中应用。（[1, 2]）来自各个国家的几篇文献报道了基于细胞的免疫疗法在各种实体瘤和血液系统恶性肿瘤中的成功应用。（[3 - 8]）我们团队与免疫细胞的关联始于我致力于RNA转录组分析以了解HIV携带者的免疫系统，这反过来需要体外扩增人类自然杀伤（NK）细胞。（[9]）这导致了方案的定制，从而成功地在体外扩增、激活NK细胞和T细胞用于免疫疗法。日本生物疗法研究所（BIJ）的目的是支持将NK细胞、γδT细胞、αβT细胞、细胞毒性T淋巴细胞（CTL）和树突状细胞（DC）等免疫细胞作为自体免疫增强疗法（AIET）用于对抗癌症的研究和临床应用。自2004年至今，BIJ已使用NK细胞、CTL、DC等进行的AIET治疗了超过5000名患者。
AIET的原理：对于使用NK细胞的AIET，该过程包括从患者外周血中分离淋巴细胞，然后使用扩增试剂盒（BINKIT，BIJ，日本）在无饲养层的情况下选择性扩增NK细胞，随后输注扩增激活的NK细胞。（[10, 11]）正如报道所示，与正常个体相比，癌症患者外周血NK细胞的活性较低（[12]），并且由于NK细胞的体外扩增可使细胞毒性能力提高5至10倍，（[13]）因此在AIET中，NK细胞先在体内扩增，然后输注给患者。我们也在致力于使用NK细胞和CTL以及NK细胞和肽脉冲DC的联合免疫疗法。联合NK细胞和CTL背后的原理是一种结合先天免疫系统和适应性免疫系统的双重优势方法，其中CTL将杀死表达MHC的癌细胞，而NK细胞也将杀死不表达MHC的癌细胞。（[10]）在NK细胞和DC的情况下，DC反过来会激活CTL，从而产生上述双重优势。我们最近提出，使用扩增的NK细胞的AIET，特别是与单克隆抗体药物联合使用时，可能是癌症免疫疗法的非常有用的工具。（[14]）
在我们对NOG SCID小鼠的研究中，活化的NK细胞显示可减小乳腺癌细胞（MDA - MB231）的大小（[15]），并且腹水的体积也抑制了胸腔积液淋巴瘤（PEL）细胞的肺转移。（[16]）在一名原发性肺腺癌患者中，AIET与热疗联合应用，CEA值从4.8 ng/ml降至1.6 ng/ml，SLX从30 U/ml降至27 U/ml，IAP从300 μg/ml降至22 μg/ml，同时肺部病变在四个月内消退。一名55岁患有乳腺浸润性导管癌并伴有肺转移的女性患者，接受曲妥珠单抗联合多次注射活化自然杀伤（NK）细胞（每隔两周一次）治疗后，肿瘤标志物下降。无进展生存期为10个月。（[11]）52例患有肺、乳腺、结肠、前列腺、肝、肾、卵巢等器官晚期癌症且对传统疗法难治性的患者，在接受热疗与基于NK细胞和CTL的免疫细胞疗法联合低剂量化疗后，52例患者中有18例观察到客观反应，包括根据实体瘤疗效评价标准（RECIST）指南评估的1例完全缓解（CR）和17例部分缓解（PR）。16例患者病情稳定（SD），而18例患者病情进展（PD）。疾病控制率为66%，包括CR、PR和SD。治疗六个月后，客观反应率和疾病控制率分别为25%和52%。这些患者均未出现不良反应。（[17]）
癌症必须通过多管齐下的方法来应对，将基于NK细胞和CTL细胞的AIET与手术、化疗和放疗等传统治疗方式以及热疗、质子束疗法和低剂量化疗等其他方式相结合，即使对于对传统治疗方式难治的晚期癌症也是有效的。
